The objective of this ShEEP proposal is to request Simoa SR-Plex Digital Immunoassay Platform (HD-1 Analyzer),which will offer researchers access to ultra-sensitive biomarker detection capabilities in a compact and affordable system. It is a digital immunoassay instrument capable of analyzing samples using the proprietary single-molecule array (Simoa) technology, and delivering ultra-sensitive measurements of proteins of interest over a wide dynamic range and with low coefficient variations (CV's). The Sioma Analyzer will aid in establishing ultra-sensitive immunoassays to serve VA-funded projects to study mechanisms, diagnosis, and treatments in various fields of study such as: inflammation, metabolism, neurology, oncology and cardiovascular disease. The projects cover an array of specialties as described below. Each Principal Investigator (PI) will benefit from Simoa Single Molecular Immunoassay Platform HD-1 Analyzer, which will establish an ultra-sensitive method for the detection and quantification of biomarkers previously difficult or impossible to measure. These applications are of great use for VA Biomedical Laboratory Research and Development Service (BLR&D) and Rehabilitation Research and Development (RR&D) funded investigators. Bingren Hu, M.D., Ph.D. will serve as the Project Director (PD)/ Principal Investigator (PI) for this proposal. Each investigator?s studies are vital to Veteran Health Care. Dr. Hu (PD/PI) will use the analyzer to test a novel hypothesis that brain ischemia- reperfusion leads to a cascade of events resulting in inactivation; as well as study novel mechanisms underlying brain ischemia-reperfusion injury and identify new therapeutic targets for the treatment of brain IR injury. Tibor Kristian, Ph.D.?s study focuses on understanding injury mechanisms that lead to mitochondrial and ultimately cellular bioenergetic failure in stoke victims. Using this new system will establish the role of cell-type-specific disruption of mitochondrial dynamics of acute brain injury in animal brain ischemia models. Bogdan Stoica, M.D. will use this new system to investigate neuro-inflammatory processes in Traumatic brain injury (TBI). TBI is a major cause of death and long-term disability in both active-duty and Veterans military populations. Sergei Atamas, M.D., Ph.D. will use this system to investigate immunue- mediated tissue fibrosis. Tissue fibrosis is a major health issues in Veterans populations. Newly funded VA investigator France Carrier, Ph.D.?s research involves delineating the role of Dual Oxidase 2 (DUOX2) as a mediator of hyper-radiosensitvity in gastric cancer. An ultra-sensitive instrument such as the Simoa HD-1 will be particularly valuable to measure low levels of protein carbonylation in mice serum following radiotherapy. Tapas Makar, Ph.D. has a VA project that studies the effects of 7, 8-dihydroxyflavone DHF, which has potent brain derived neurotrophic factor (BDNF) agonist activity that can be given orally and readily crosses the blood brain barrier as a potential treatment in a model of Multiple sclerosis (MS). This is currently being studied in a murine model of MS, and Dr. Makar will use the machine?s ultrasensitive assays for his award. Finally, Leonardo Tonelli, Ph.D.?s project studies the stress of deployment and exposure to traumatic events that puts soldiers at a greater risk than the general public for the development of psychological disorders- including anxiety and depression, as well as post-traumatic stress disorder. He will use the analyzer to test preliminary results that strongly indicate that cluster of differentiation 8 positive (CD8+) cytotoxic T cells are a major source of systemic and brain inflammation, and promote susceptibility to develop maladaptive behavioral responses to stress.